Bio-On-Magnetic-Beads (BOMB): Open platform for high-throughput nucleic acid extraction and manipulation

PLoS Biol. 2019 Jan 10;17(1):e3000107. doi: 10.1371/journal.pbio.3000107. eCollection 2019 Jan.

Abstract

Current molecular biology laboratories rely heavily on the purification and manipulation of nucleic acids. Yet, commonly used centrifuge- and column-based protocols require specialised equipment, often use toxic reagents, and are not economically scalable or practical to use in a high-throughput manner. Although it has been known for some time that magnetic beads can provide an elegant answer to these issues, the development of open-source protocols based on beads has been limited. In this article, we provide step-by-step instructions for an easy synthesis of functionalised magnetic beads, and detailed protocols for their use in the high-throughput purification of plasmids, genomic DNA, RNA and total nucleic acid (TNA) from a range of bacterial, animal, plant, environmental and synthetic sources. We also provide a bead-based protocol for bisulfite conversion and size selection of DNA and RNA fragments. Comparison to other methods highlights the capability, versatility, and extreme cost-effectiveness of using magnetic beads. These open-source protocols and the associated webpage (https://bomb.bio) can serve as a platform for further protocol customisation and community engagement.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • DNA / isolation & purification
  • High-Throughput Nucleotide Sequencing / methods*
  • Humans
  • Magnetic Fields
  • Microspheres
  • Nucleic Acids / isolation & purification*
  • RNA / isolation & purification

Substances

  • Nucleic Acids
  • RNA
  • DNA

Grant support

This work was supported by the University of Otago, Department of Anatomy (TH) and Deutsche Forschungsgemeinschaft—www.dfg.de (DFG SPP1784) (TPJ). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.